Camptothecin derivatives and uses thereof

ABSTRACT

Provided herein are camptothecin derivatives and pharmaceutical compositions thereof. In other embodiments, provided herein are methods of treatment, prevention, or amelioration of a variety of medical disorders such as, for example, cancer using the compounds and pharmaceutical compositions disclosed herein.

This application claims priority under 35 U.S.C. § 119 (e) from U.S.Provisional Application Ser. No. 62/305,519 filed Mar. 8, 2016, which ishereby incorporated by reference in its entirety.

FIELD

Provided herein are camptothecin derivatives and pharmaceuticalcompositions thereof. In other embodiments, provided herein are methodsof treatment, prevention, or amelioration of a variety of medicaldisorders such as, for example, cancer using the compounds andpharmaceutical compositions disclosed herein.

BACKGROUND

Camptothecin (CPT), originally isolated from the Chinese treeCamptotheca acuminate, possesses potent antitumor properties due to itsinhibition of topoisomerase I (Hsiang et al., J Biol Chem 1985, 260,4873; Layergne et al., Cancer Res 1999, 59, 2939; Saltz et al., Lancet2003, 361, 2235). However, CPT exhibits several properties that severelylimit its clinical application such as low aqueous solubility, (Lerchenet al., J Med Chem 2001, 44, 4186; Klein et al., J Med Chem 2003, 46,190; Saltz et al., Id.) high levels of protein binding and rapidinactivation through lactone ring hydrolysis. The insolubility of CPThas severely restricted its clinical application and has led to thedevelopment of several water-soluble congeners, which are in variousphases of clinical trials or in the clinic (Potmesil, Cancer Res 1994,54, 1431). Camptothecin undergoes a reversible, pH-dependent ringopening reaction between the active lactone (closed E-ring) and inactivecarboxylate (open E-ring) form, which has also been shown to be toxic:Lactone hydrolysis is also enhanced by the specific binding andsequestration of the carboxylate form to various proteins, such as humanserum albumin, in the biological matrix, which shifts the equilibriumfurther toward the carboxylate form. Thus, the clinical utility of CPThas been severely hindered by the hydrolytic instability of itsE-lactone ring in blood serum. Additionally, considerable variability inthe oral and intravenous bioavailability of CPT suggests poor cellularand tumor uptake of unmodified CPT drugs (Sinka et al., J ControlRelease 2004, 100, 275).

Structure of Camptothecin (CPT) in Lactone (1) and Carboxylate (2) Forms

The issues, above, are not limited to camptothecin but are also sharedto certain extent by many of the pharmacologically active derivatives ofcamptothecin. Accordingly, what are needed are new compositions of newanalogs of camptothecin and camptothecin derivatives that allow greaterstability and delivery of active drugs, while maintaining high loadingand concentrations of the drug in the composition.

SUMMARY

Provided herein are camptothecin derivatives which address these andother needs. In one aspect, a compound of Formula (I) is provided:

or salts, hydrates or solvates thereof, wherein:

R₁ is hydrogen, alkyl, substituted alkyl, heteroalkyl, substitutedheteroalkyl, —SiR₆R₇R₈, alkyl substituted with one or more SiR₆R₇R₈,—CH═NOR₁₃ or optionally R₁ and R₂ together with the atom to which theyare connected form a 5, 6 or 7 membered cycloalkyl or cycloheteroalkylring or substituted cycloalkyl or cycloheteroalkyl ring;

R₂ is hydrogen, alkyl, substituted alkyl —NO₂ or —NH₂;

R₃ is hydrogen, alkyl, substituted alkyl, acyl, substituted acyl, or—OR₉ or optionally R₃ and R₄ together with the atom to which they areconnected form a 5, 6 or 7 membered cycloalkyl or cycloheteroalkyl ringor substituted cycloalkyl or cycloheteroalkyl ring

R₄ is hydrogen or halogen;

R₅ is alkyl, aryl, arylalkyl, acyl or -(L)_(n)-P;

R₆, R₇ or R₈ are independently alkyl or arylalkyl;

R₉ is hydrogen, alkyl, substituted alkyl, —C(O)R₁₀—C(O)OR₁₀ or—C(O)NR₁₁R₁₂;

R₁₀ is alkyl, substituted alkyl, arylalkyl, substituted alkyl,heteroalkyl or substituted heteroalkyl;

R₁₁, R₁₂ and R₁₃ are independently alkyl, substituted alkyl, arylalkyl,substituted alkyl, heteroalkyl or substituted heteroalkyl or optionallyR₁₂ and R₁₃ together with the atom to which they are connected form a 5,6 or 7 membered cycloheteroalkyl ring or substituted cycloheteroalkylring;

L is a linker;

P is a polymer; and

n is 0 or 1;

provided that when R₁, R₂, R₃ and R₄ are hydrogen that R₅ is not -Ph,—C₂H₅, —C₅H₁₁, —C₆H₁₃, —C₇H₁₅, —C₈H₁₇, —C₁₅H₃₁, —C₃H₆C≡CH, —C₃H₆C≡CH,—C₃H₆C═CH₂ or —C═CH(CH₃);

provided that when R₁, R₃ and R₄ are hydrogen and R₂, is —NO₂ that R₅ isnot -Ph or —C₅H₁₁;

provided that when R₁, R₃ and R₄ are hydrogen, and R₂, is —NH₂ that R₅is not -Ph, —C₅H₁₁ or —C₈H₁₇;

provided that when R₁, R₂ and R₄ are hydrogen and R₃ is —OCH₃ that R₅ isnot —C₂H₅ or —C₅H₁₁;

provided that when R₂, R₃ and R₄ are hydrogen and R₁ is —CH₃ that R₅ isnot -Ph or —C₆H₁₃;

provided that when R₂, R₃ and R₄ are hydrogen and R₁ is —CH₂OH that R₅is not —C₅H₁₁ or —C₆H₁₃;

provided that when R₂, R₃ and R₄ are hydrogen and R₁

is that R₅ is not —C₆H₁₃;

provided that when R₂, R₃ and R₄ are hydrogen and R₁

is that R₅ is not —C₅H₁₁;

provided that when R₂, R₃ and R₄ are hydrogen and R₁

is that R₅ is not -Ph;

provided that when R₁, R₂ and R₄ are hydrogen and R₃ is —C(O)R₁₀ that R₅and R₁₀ are not identical;

provided that when R₂ and R₄ are hydrogen, R₁ is —C₂H₅ and R₃ is—C(O)R₁₀ that R₅ and R₁₀ are not identical;

provided that when R₂ and R₄ are hydrogen, R₁ is —C₂H₅ and R₃ is —H thatR₅ is not C₁₀H₂₁; and

provided that when R₂ and R₄ are hydrogen, R₁ is —C₂H₅ and R₃ is—C(O)OtBu that R₅ is not C₁₀H₂₁.

Also provided are derivatives, including salts, esters, enol ethers,enol esters, solvates, hydrates and metabolites of the compoundsdescribed herein. Further provided are pharmaceutical compositions whichinclude the compounds provided herein and a vehicle.

Methods of treating, preventing, or ameliorating symptoms of medicaldisorders such as, for example, cancer or infection, are also providedherein. In practicing the methods, therapeutically effective amounts ofthe compounds or pharmaceutical compositions thereof are administered toa subject.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates the percentage reduction of Alamar Blue versus cellnumber of SW620 cells over a 1, 2, 4, 7 and 24 hour period;

FIG. 2 illustrates the percentage reduction of Alamar Blue versus cellnumber of CT26 cells over a 1, 2, 4, 7 and 24 hour period;

FIG. 3 illustrates the percentage inhibition of HT29 cells exposed to 2,3 and SN38;

FIG. 4 illustrates the percentage inhibition of SW620 cells exposed to2, 3 and SN38; and

FIG. 5 illustrates the percentage inhibition of CT26 cells exposed to 2,3 and CT26.

DETAILED DESCRIPTION Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of ordinary skillin the art to which this invention belongs. In the event that there is aplurality of definitions for a term herein, those in this sectionprevail unless stated otherwise.

“Alkyl” by itself or as part of another substituent, refers to asaturated or unsaturated, branched, straight-chain or cyclic monovalenthydrocarbon radical derived by the removal of one hydrogen atom from asingle carbon atom of a parent alkane, alkene or alkyne. Typical alkylgroups include, but are not limited to, methyl; ethyls such as ethanyl,ethenyl, ethynyl; propyls such as propan-1-yl, propan-2-yl,cyclopropan-1-yl, prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl(allyl), cycloprop-1-en-1-yl; cycloprop-2-en-1-yl, prop-1-yn-1-yl,prop-2-yn-1-yl, etc.; butyls such as butan-1-yl, butan-2-yl,2-methyl-propan-1-yl, 2-methyl-propan-2-yl, cyclobutan-1-yl,but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl,but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl,cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl,but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like. Theterm “alkyl” is specifically intended to include groups having anydegree or level of saturation, i.e., groups having exclusively singlecarbon-carbon bonds, groups having one or more double carbon-carbonbonds, groups having one or more triple carbon-carbon bonds and groupshaving mixtures of single, double and triple carbon-carbon bonds. Wherea specific level of saturation is intended, the expressions “alkanyl,”“alkenyl,” and “alkynyl” are used. In some embodiments, an alkyl groupcomprises from 1 to 20 carbon atoms (C₁-C₂₀ alkyl). In otherembodiments, an alkyl group comprises from 1 to 10 carbon atoms (C₁-C₁₀alkyl). In still other embodiments, an alkyl group comprises from 1 to 6carbon atoms (C₁-C₆ alkyl).

“Alkanyl” by itself or as part of another substituent, refers to asaturated branched, straight-chain or cyclic alkyl radical derived bythe removal of one hydrogen atom from a single carbon atom of a parentalkane. Typical alkanyl groups include, but are not limited to,methanyl; ethanyl; propanyls such as propan-1-yl, propan-2-yl(isopropyl), cyclopropan-1-yl, etc.; butanyls such as butan-1-yl,butan-2-yl (sec-butyl), 2-methyl-propan-1-yl (isobutyl),2-methyl-propan-2-yl (t-butyl), cyclobutan-1-yl, etc.; and the like.

“Alkenyl” by itself or as part of another substituent, refers to anunsaturated branched, straight-chain or cyclic alkyl radical having atleast one carbon-carbon double bond derived by the removal of onehydrogen atom from a single carbon atom of a parent alkene. The groupmay be in either the cis or trans conformation about the double bond(s).Typical alkenyl groups include, but are not limited to, ethenyl;propenyls such as prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl(allyl), prop-2-en-2-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl;butenyls such as but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl,but-2-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, buta-1,3-dien-1-yl,buta-1,3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl,cyclobuta-1,3-dien-1-yl, etc.; and the like.

“Alkynyl” by itself or as part of another substituent refers to anunsaturated branched, straight-chain or cyclic alkyl radical having atleast one carbon-carbon triple bond derived by the removal of onehydrogen atom from a single carbon atom of a parent alkyne. Typicalalkynyl groups include, but are not limited to, ethynyl; propynyls suchas prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butynyls such as but-1-yn-1-yl,but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like.

“Acyl” by itself or as part of another substituent refers to a radical—C(O)R⁴⁰⁰, where R⁴⁰⁰ is hydrogen, alkyl, substituted alkyl, aryl,substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl,substituted heteroalkyl, heteroarylalkyl or substituted heteroarylalkylas defined herein. Representative examples include, but are not limitedto formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl,benzoyl, benzylcarbonyl and the like.

“Aryl” by itself or as part of another substituent, refers to amonovalent aromatic hydrocarbon group derived by the removal of onehydrogen atom from a single carbon atom of a parent aromatic ringsystem, as defined herein. Typical aryl groups include, but are notlimited to, groups derived from aceanthrylene, acenaphthylene,acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene,fluoranthene, fluorene, hexacene, hexaphene, hexalene, as-indacene,s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene,ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene,phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene,rubicene, triphenylene, trinaphthalene and the like. In someembodiments, an aryl group comprises from 6 to 20 carbon atoms (C₆-C₂₀aryl). In other embodiments, an aryl group comprises from 6 to 15 carbonatoms (C₆-C₁₅ aryl). In still other embodiments, an aryl group comprisesfrom 6 to 15 carbon atoms (C₆-C₁₀ aryl).

“Arylalkyl” by itself or as part of another substituent, refers to anacyclic alkyl group in which one of the hydrogen atoms bonded to acarbon atom, typically a terminal or sp³ carbon atom, is replaced withan aryl group as, as defined herein. Typical arylalkyl groups include,but are not limited to, benzyl, 2-phenylethan-1-yl, 2-phenylethen-1-yl,naphthylmethyl, 2-naphthylethan-1-yl, 2-naphthylethen-1-yl,naphthobenzyl, 2-naphthophenylethan-1-yl and the like. Where specificalkyl moieties are intended, the nomenclature arylalkanyl, arylalkenyland/or arylalkynyl is used. In some embodiments, an arylalkyl group is(C₆-C₃₀) arylalkyl, e.g., the alkanyl, alkenyl or alkynyl moiety of thearylalkyl group is (C₁-C₁₀) alkyl and the aryl moiety is (C₆-C₂₀) aryl.In other embodiments, an arylalkyl group is (C₆-C₂₀) arylalkyl, e.g.,the alkanyl, alkenyl or alkynyl moiety of the arylalkyl group is (C₁-C₈)alkyl and the aryl moiety is (C₆-C₁₂) aryl. In still other embodiments,an arylalkyl group is (C₆-C₁₅) arylalkyl, e.g., the alkanyl, alkenyl oralkynyl moiety of the arylalkyl group is (C₁-C₅) alkyl and the arylmoiety is (C₆-C₁₀) aryl.

“Compounds” refers to compounds encompassed by structural formulaedisclosed herein and includes any specific compounds within theseformulae whose structure is disclosed herein. Compounds may beidentified either by their chemical structure and/or chemical name. Whenthe chemical structure and chemical name conflict, the chemicalstructure is determinative of the identity of the compound. Thecompounds described herein may contain one or more chiral centers and/ordouble bonds and therefore, may exist as stereoisomers, such asdouble-bond isomers (i.e., geometric isomers), enantiomers ordiastereomers. Accordingly, the chemical structures depicted hereinencompass all possible enantiomers and stereoisomers of the illustratedcompounds including the stereoisomerically pure form (e.g.,geometrically pure, enantiomerically pure or diastereomerically pure)and enantiomeric and stereoisomeric mixtures. Enantiomeric andstereoisomeric mixtures can be resolved into their component enantiomersor stereoisomers using separation techniques or chiral synthesistechniques well known to the skilled artisan. The compounds may alsoexist in several tautomeric forms including the enol form, the keto formand mixtures thereof. Accordingly, the chemical structures depictedherein encompass all possible tautomeric forms of the illustratedcompounds. The compounds described also include isotopically labeledcompounds where one or more atoms have an atomic mass different from theatomic mass conventionally found in nature. Examples of isotopes thatmay be incorporated into the compounds described herein include, but arenot limited to, ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³⁵S, etc. In general,it should be understood that all isotopes of any of the elementscomprising the compounds described herein may be found in thesecompounds. Compounds may exist in unsolvated or unhydrated forms as wellas solvated forms, including hydrated forms and as N-oxides. In general,compounds may be hydrated, solvated or N-oxides. Certain compounds mayexist in multiple crystalline or amorphous forms. In general, allphysical forms are equivalent for the uses contemplated herein and areintended to be within the scope of the present invention. Further, itshould be understood, when partial structures of the compounds areillustrated, that brackets indicate the point of attachment of thepartial structure to the rest of the molecule.

“Heteroalkyl,” “Heteroalkanyl,” “Heteroalkenyl” and “Heteroalkynyl” bythemselves or as part of other substituents, refer to alkyl, alkanyl,alkenyl and alkynyl groups, respectively, in which one or more of thecarbon atoms (and optionally any associated hydrogen atoms), are each,independently of one another, replaced with the same or differentheteroatoms or heteroatomic groups. Typical heteroatoms or heteroatomicgroups which can replace the carbon atoms include, but are not limitedto, —O—, —S—, —N—, —Si—, —NH—, —S(O)—, —S(O)₂—, —S(O)NH—, —S(O)₂NH— andthe like and combinations thereof. The heteroatoms or heteroatomicgroups may be placed at any interior position of the alkyl, alkenyl oralkynyl groups. Typical heteroatomic groups which can be included inthese groups include, but are not limited to, —O—, —S—, —O—O—, —S—S—,—O—S—, —NR⁵⁰¹R⁵⁰²—, ═N—N═, —N═N—, —N═N—NR⁵⁰³R⁵⁰⁴, —PR⁵⁰⁵—, —P(O)₂—,—POR⁵⁰⁶—, —O—P(O)₂—, —SO—, —SO₂—, —SnR⁵⁰⁷R⁵⁰⁸— and the like, where R⁵⁰¹,R⁵⁰², R⁵⁰³, R⁵⁰⁴, R⁵⁰⁵, R⁵⁰⁶, R⁵⁰⁷ and R⁵⁰⁸ are independently hydrogen,alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl,substituted cycloheteroalkyl, heteroalkyl, substituted heteroalkyl,heteroaryl, substituted heteroaryl, heteroarylalkyl or substitutedheteroarylalkyl.

“Heteroaryl” by itself or as part of another substituent, refers to amonovalent heteroaromatic radical derived by the removal of one hydrogenatom from a single atom of a parent heteroaromatic ring systems, asdefined herein. Typical heteroaryl groups include, but are not limitedto, groups derived from acridine, β-carboline, chromane, chromene,cinnoline, furan, imidazole, indazole, indole, indoline, indolizine,isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline,isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, perimidine,phenanthridine, phenanthroline, phenazine, phthalazine, pteridine,purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine,pyrrole, pyrrolizine, quinazoline, quinoline, quinolizine, quinoxaline,tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene, and thelike. In some embodiments, the heteroaryl group comprises from 5 to 20ring atoms (5-20 membered heteroaryl). In other embodiments, theheteroaryl group comprises from 5 to 10 ring atoms (5-10 memberedheteroaryl). Exemplary heteroaryl groups include those derived fromfuran, thiophene, pyrrole, benzothiophene, benzofuran, benzimidazole,indole, pyridine, pyrazole, quinoline, imidazole, oxazole, isoxazole andpyrazine.

“Heteroarylalkyl” by itself or as part of another substituent refers toan acyclic alkyl group in which one of the hydrogen atoms bonded to acarbon atom, typically a terminal or sp³ carbon atom, is replaced with aheteroaryl group. Where specific alkyl moieties are intended, thenomenclature heteroarylalkanyl, heteroarylalkenyl and/orheteroarylalkynyl is used. In some embodiments, the heteroarylalkylgroup is a 6-21 membered heteroarylalkyl, e.g., the alkanyl, alkenyl oralkynyl moiety of the heteroarylalkyl is (C₁-C₆) alkyl and theheteroaryl moiety is a 5-15-membered heteroaryl. In other embodiments,the heteroarylalkyl is a 6-13 membered heteroarylalkyl, e.g., thealkanyl, alkenyl or alkynyl moiety is (C₁-C₃) alkyl and the heteroarylmoiety is a 5-10 membered heteroaryl.

“Hydrates” refers to incorporation of water into to the crystal latticeof a compound described herein, in stoichiometric proportions, resultingin the formation of an adduct. Methods of making hydrates include, butare not limited to, storage in an atmosphere containing water vapor,dosage forms that include water, or routine pharmaceutical processingsteps such as, for example, crystallization (i.e., from water or mixedaqueous solvents), lyophilization, wet granulation, aqueous filmcoating, or spray drying. Hydrates may also be formed, under certaincircumstances, from crystalline solvates upon exposure to water vapor,or upon suspension of the anhydrous material in water. Hydrates may alsocrystallize in more than one form resulting in hydrate polymorphism. Seee.g., (Guillory, K., Chapter 5, pp. 202-205 in Polymorphism inPharmaceutical Solids, (Brittain, H. ed.), Marcel Dekker, Inc., NewYork, N.Y., 1999). The above methods for preparing hydrates are wellwithin the ambit of those of skill in the art, are completelyconventional and do not require any experimentation beyond what istypical in the art. Hydrates may be characterized and/or analyzed bymethods well known to those of skill in the art such as, for example,single crystal X-ray diffraction, X-ray powder diffraction, Polarizingoptical microscopy, thermal microscopy, thermogravimetry, differentialthermal analysis, differential scanning calorimetry, IR spectroscopy,Raman spectroscopy and NMR spectroscopy. (Brittain, H., Chapter 6, pp.205-208 in Polymorphism in Pharmaceutical Solids, (Brittain, H. ed.),Marcel Dekker, Inc. New York, 1999). In addition, many commercialcompanies routine offer services that include preparation and/orcharacterization of hydrates such as, for example, HOLODIAG, PharmaparcII, Voie de l'Innovation, 27 100 Val de Reuil, France(http://www.holodiag.com).

“Parent Aromatic Ring System” refers to an unsaturated cyclic orpolycyclic ring system having a conjugated π electron system.Specifically included within the definition of “parent aromatic ringsystem” are fused ring systems in which one or more of the rings arearomatic and one or more of the rings are saturated or unsaturated, suchas, for example, fluorene, indane, indene, phenalene, etc. Typicalparent aromatic ring systems include, but are not limited to,aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene,benzene, chrysene, coronene, fluoranthene, fluorene, hexacene,hexaphene, hexalene, as-indacene, s-indacene, indane, indene,naphthalene, octacene, octaphene, octalene, ovalene, penta-2,4-diene,pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene,picene, pleiadene, pyrene, pyranthrene, rubicene, triphenylene,trinaphthalene and the like.

“Preventing” or “prevention” refers to a reduction in risk of acquiringa disease or disorder (i.e., causing at least one of the clinicalsymptoms of the disease not to develop in a patient that may be exposedto or predisposed to the disease but does not yet experience or displaysymptoms of the disease). In some embodiments, “preventing” or“prevention” refers to reducing symptoms of the disease by taking thecompound in a preventative fashion. The application of a therapeutic forpreventing or prevention of a disease of disorder is known as‘prophylaxis.’ In some embodiments, the compounds provided hereinprovide superior prophylaxis because of lower long term side effectsover long time periods.

“Prodrug” refers to a derivative of a drug molecule that requires atransformation within the body to release the active drug. Prodrugs arefrequently (though not necessarily) pharmacologically inactive untilconverted to the parent drug.

“Promoiety” refers to a form of protecting group that when used to maska functional group within a drug molecule converts the drug into aprodrug. Typically, the promoiety will be attached to the drug viabond(s) that are cleaved by enzymatic or non-enzymatic means in vivo.

“Salt” refers to a salt of a compound, which possesses the desiredpharmacological activity of the parent compound. Such salts include: (1)acid addition salts, formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andthe like; or formed with organic acids such as acetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound isreplaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, N-methylglucamine and thelike. In some embodiments, the salt is pharmaceutically acceptable.

“Solvates” refers to incorporation of solvents into to the crystallattice of a compound described herein, in stoichiometric proportions,resulting in the formation of an adduct. Methods of making solvatesinclude, but are not limited to, storage in an atmosphere containing asolvent, dosage forms that include the solvent, or routinepharmaceutical processing steps such as, for example, crystallization(i.e., from solvent or mixed solvents) vapor diffusion, etc. Solvatesmay also be formed, under certain circumstances, from other crystallinesolvates or hydrates upon exposure to the solvent or upon suspensionmaterial in solvent. Solvates may crystallize in more than one formresulting in solvate polymorphism. See e.g., (Guillory, K., Chapter 5,pp. 205-208 in Polymorphism in Pharmaceutical Solids, (Brittain, H.ed.), Marcel Dekker, Inc., New York, N.Y., 1999)). The above methods forpreparing solvates are well within the ambit of those of skill in theart, are completely conventional do not require any experimentationbeyond what is typical in the art. Solvates may be characterized and/oranalyzed by methods well known to those of skill in the art such as, forexample, single crystal X-ray diffraction, X-ray powder diffraction,Polarizing optical microscopy, thermal microscopy, thermogravimetry,differential thermal analysis, differential scanning calorimetry, IRspectroscopy, Raman spectroscopy and NMR spectroscopy. (Brittain, H.,Chapter 6, pp. 205-208 in Polymorphism in Pharmaceutical Solids,(Brittain, H. ed.), Marcel Dekker, Inc. New York, 1999). In addition,many commercial companies routine offer services that includepreparation and/or characterization of solvates such as, for example,HOLODIAG, Pharmaparc II, Voie de l'Innovation, 27 100 Val de Reuil,France (http://www.holodiag.com).

“Substituted” when used to modify a specified group or radical, meansthat one or more hydrogen atoms of the specified group or radical areeach, independently of one another, replaced with the same or differentsubstituent(s). Substituent groups useful for substituting saturatedcarbon atoms in the specified group or radical include, but are notlimited to —R^(a), halo, —O⁻, ═O, —OR^(b), —SR^(b), —S⁻, ═S,—NR^(c)R^(c), ═NR^(b), ═N—OR^(b), trihalomethyl, —CF₃, —CN, —OCN, —SCN,—NO, —NO₂, ═N₂, —N₃, —S(O)₂R^(b), —S(O)₂NR^(b), —S(O)₂O⁻, —S(O)₂OR^(b),—OS(O)₂R^(b), —OS(O)₂O⁻, —OS(O)₂OR^(b), —P(O)(O⁻)₂, —P(O)(OR^(b))(O⁻),—P(O)(OR^(b))(OR^(b)), —C(O)R^(b), —C(S)R^(b), —C(NR^(b))R^(b), —C(O)O⁻,—C(O)OR^(b), —C(S)OR^(b), —C(O)NR^(c)R^(c), —C(NR^(b))NR^(c)R^(c),—OC(O)R^(b), —OC(S)R^(b), —OC(O)O⁻, —OC(O)OR^(b), —OC(S)OR^(b),—NR^(b)C(O)R^(b), —NRfC(S)R^(b), —NR^(b)C(O)O⁻, —NR^(b)C(O)OR^(b),—NR^(b)C(S)OR^(b), —NR^(b)C(O)NR^(c)R^(c), —NR^(b)C(NR^(b))R^(b) and—NR^(b)C(NR^(b))NR^(c)R^(c), where R^(a) is selected from the groupconsisting of alkyl, cycloalkyl, heteroalkyl, cycloheteroalkyl, aryl,arylalkyl, heteroaryl and heteroarylalkyl; each R^(b) is independentlyhydrogen or R^(a); and each R^(c) is independently R^(b) oralternatively, the two R^(c)'s are taken together with the nitrogen atomto which they are bonded form a 4-, 5-, 6- or 7-memberedcycloheteroalkyl which may optionally include from 1 to 4 of the same ordifferent additional heteroatoms selected from the group consisting ofO, N and S. As specific examples, —NR^(c)R^(c) is meant to include —NH₂,—NH-alkyl, N-pyrrolidinyl and N-morpholinyl.

Similarly, substituent groups useful for substituting unsaturated carbonatoms in the specified group or radical include, but are not limited to,—R^(a), halo, —O—, —OR^(b), —SR^(b), —S⁻, —NR^(c)R^(c), trihalomethyl,—CF₃, —CN, —OCN, —SCN, —NO, —NO₂, —N₃, —S(O)₂R^(b), —S(O)₂O⁻,—S(O)₂OR^(b), —OS(O)₂R^(b), —OS(O)₂O⁻, —OS(O)₂OR^(b), —P(O)(O⁻)₂,—P(O)(OR^(b)(O⁻), —P(O)(OR^(b))(OR^(b)), —C(O)R^(b), —C(S)R^(b),—C(NR^(b))R^(b), —C(O)O⁻, —C(O)OR^(b), —C(S)OR^(b), —C(O)NR^(c)R^(c),—C(NR^(b))NR^(c)R^(c), —OC(O)R^(b), —OC(S)R^(b), —OC(O)O⁻, —OC(O)OR^(b),—OC(S)OR^(b), —NR^(b)C(O)R^(b), —NR^(b)C(S)R^(b), —NR^(b)C(O)O⁻,—NR^(b)C(O)OR^(b), —NR^(b)C(S)OR^(b), —NR^(b)C(O)NR^(c)R^(c),—NR^(b)C(NR^(b))R^(b) and —NR^(b)C(NR^(b))NR^(c)R^(c), where R^(a),R^(b) and R^(c) are as previously defined.

Substituent groups useful for substituting nitrogen atoms in heteroalkyland cycloheteroalkyl groups include, but are not limited to, —R^(a),—O—, —OR^(b), —SR^(b), —S⁻, —NR^(c)R^(c), trihalomethyl, —CF₃, —CN, —NO,—NO₂, —S(O)₂R^(b), —S(O)₂O⁻, —S(O)₂OR^(b), —OS(O)₂R^(b), —OS(O)₂O⁻,—OS(O)₂OR^(b), —P(O)(O⁻)₂, —P(O)(OR^(b))(O⁻), —P(O)(OR^(b))(OR^(b)),—C(O)R^(b), —C(S)R^(b), —C(NR^(b))R^(b), —C(O)OR^(b), —C(S)OR^(b),—C(O)NR^(c)R^(c), —C(NR^(b))NR^(c)R^(c), —OC(O)R^(b), —OC(S)R^(b),—OC(O)OR^(b), —OC(S)OR^(b), —NR^(b)C(O)R^(b), —NR^(b)C(S)R^(b),—NR^(b)C(O)OR^(b), —NR^(b)C(S)OR^(b), —NR^(b)C(O)NR^(c)R^(c),—NR^(b)C(NR^(b))R^(b) and —NR^(b)C(NR^(b))NR^(c)R^(c), where R^(a),R^(b) and R^(c) are as previously defined.

Substituent groups from the above lists useful for substituting otherspecified groups or atoms will be apparent to those of skill in the art.The substituents used to substitute a specified group can be furthersubstituted, typically with one or more of the same or different groupsselected from the various groups specified above. In some embodiments,substituents are limited to the groups above.

“Subject,” “individual” or “patient” is used interchangeably herein andrefers to a vertebrate, preferably a mammal. Mammals include, but arenot limited to, murines, rodents, simians, humans, farm animals, sportanimals and pets.

“Treating” or “treatment” of any disease or disorder refers, in someembodiments, to ameliorating the disease or disorder (i.e., arresting orreducing the development of the disease or at least one of the clinicalsymptoms thereof,). Treatment may also be considered to includepreemptive or prophylactic administration to ameliorate, arrest orprevent the development of the disease or at least one of the clinicalsymptoms. Treatment can also refer to the lessening of the severityand/or the duration of one or more symptoms of a disease or disorder. Ina further feature, the treatment rendered has lower potential for longterm side effects over multiple years. In other embodiments “treating”or “treatment” refers to ameliorating at least one physical parameter,which may not be discernible by the patient. In yet other embodiments,“treating” or “treatment” refers to inhibiting the disease or disorder,either physically, (e.g., stabilization of a discernible symptom),physiologically, (e.g., stabilization of a physical parameter) or both.In yet other embodiments, “treating” or “treatment” refers to delayingthe onset of the disease or disorder.

“Therapeutically effective amount” means the amount of a compound that,when administered to a patient for treating a disease, is sufficient toeffect such treatment for the disease. The “therapeutically effectiveamount” will vary depending on the compound, the disease and itsseverity and the age, weight, adsorption, distribution, metabolism andexcretion etc., of the patient to be treated.

“Vehicle” refers to a diluent, excipient or carrier with which acompound is administered to a subject. In some embodiments, the vehicleis pharmaceutically acceptable.

Compounds

Provided herein, in one aspect, are compounds of Formula (I):

or salts, hydrates or solvates thereof wherein:

R₁ is hydrogen, alkyl, substituted alkyl, heteroalkyl, substitutedheteroalkyl, —SiR₆R₇R₈, alkyl substituted with one or more SiR₆R₇R₈,—CH═NOR₁₃ or optionally R₁ and R₂ together with the atom to which theyare connected form a 5, 6 or 7 membered cycloalkyl or cycloheteroalkylring or substituted cycloalkyl or cycloheteroalkyl ring;

R₂ is hydrogen, alkyl, substituted alkyl —NO₂ or —NH₂;

R₃ is hydrogen, alkyl, substituted alkyl, acyl, substituted acyl, or—OR₉ or optionally R₃ and R₄ together with the atom to which they areconnected form a 5, 6 or 7 membered cycloalkyl or cycloheteroalkyl ringor substituted cycloalkyl or cycloheteroalkyl ring

R₄ is hydrogen or halogen;

R₅ is alkyl, aryl, arylalkyl, acyl or -(L)_(n)-P;

R₆, R₇ or R₈ are independently alkyl or arylalkyl;

R₉ is hydrogen, alkyl, substituted alkyl, —C(O)R₁₀—C(O)OR₁₀ or—C(O)NR₁₁R₁₂;

R₁₀ is alkyl, substituted alkyl, arylalkyl, substituted alkyl,heteroalkyl or substituted heteroalkyl;

R₁₁, R₁₂ and R₁₃ are independently alkyl, substituted alkyl, arylalkyl,substituted alkyl, heteroalkyl or substituted heteroalkyl or optionallyR₁₂ and R₁₃ together with the atom to which they are connected form a 5,6 or 7 membered cycloheteroalkyl ring or substituted cycloheteroalkylring;

L is a linker;

P is a polymer; and

n is 0 or 1

provided that when R₁, R₂, R₃ and R₄ are hydrogen that R₅ is not -Ph,—C₂H₅, —C₅H₁₁, —C₆H₁₃, —C₇H₁₅, —C₈H₁₇, —C₁₅H₃₁, —C₃H₆C≡CH, —C₃H₆C≡CH,—C₃H₆C═CH₂ or —C═CH(CH₃);

provided that when R₁, R₃ and R₄ are hydrogen and R₂, is —NO₂ that R₅ isnot -Ph or —C₅H₁₁;

provided that when R₁, R₃ and R₄ are hydrogen, and R₂, is —NH₂ that R₅is not -Ph, —C₅H₁₁ or —C₈H₁₇;

provided that when R₁, R₂ and R₄ are hydrogen and R₃ is —OCH₃ that R₅ isnot —C₂H₅ or —C₅H₁₁;

provided that when R₂, R₃ and R₄ are hydrogen and R₁ is —CH₃ that R₅ isnot -Ph or —C₆H₁₃;

provided that when R₂, R₃ and R₄ are hydrogen and R₁ is —CH₂OH that R₅is not —C₅H₁₁ or —C₆H₁₃;

provided that when R₂, R₃ and R₄ are hydrogen and R₁

is that R₅ is not —C₆H₁₃;

provided that when R₂, R₃ and R₄ are hydrogen and R₁

is that R₅ is not —C₅H₁₁;

provided that when R₂, R₃ and R₄ are hydrogen and R₁

is that R₅ is not -Ph;

provided that when R₁, R₂ and R₄ are hydrogen and R₃ is —C(O)R₁₀ that R₅and R₁₀ are not identical;

provided that when R₂ and R₄ are hydrogen, R₁ is —C₂H₅ and R₃ is—C(O)R₁₀ that R₅ and R₁₀ are not identical;

provided that when R₂ and R₄ are hydrogen, R₁ is —C₂H₅ and R₃ is —H thatR₅ is not C₁₀H₂₁; and

provided that when R₂ and R₄ are hydrogen, R₁ is —C₂H₅ and R₃ is—C(O)OtBu that R₅ is not C₁₀H₂₁.

In some embodiments, a compound having the structure of Formula (II) isprovided:

In other embodiments, a compound having the structure of Formula (III)is provided:

In still other embodiments, a compound having the structure of Formula(IV) is provided:

In still other embodiments, a compound having the structure of Formula(V) is provided:

In still other embodiments, a compound having the structure of Formula(VI) is provided:

In still other embodiments, a compound having the structure of Formula(VII) is provided:

In still other embodiments, a compound having the structure of Formula(VIII) is provided:

In still other embodiments, a compound having the structure of Formula(IX) is provided:

In still other embodiments, a compound having the structure of Formula(X) is provided:

In still other embodiments, a compound having the structure of Formula(XI) is provided:

In still other embodiments, a compound having the structure of Formula(XII) is provided:

In still other embodiments, a compound having the structure of Formula(XIII) is provided:

In still other embodiments, a compound having the structure of Formula(XIV) is provided:

In some embodiments, R₁, R₂, R₃ and R₄ are not hydrogen. In otherembodiments, R₅ is alkyl or acyl. In still other embodiments, R₅ is(C₁₀-C₂₀) alkyl. In still other embodiments, R₅ is n-C₁₀H₂₁, n-C₁₁H₂₃,n-C₁₀H₂₁, n-C₁₂H₂₅, n-C₁₃H₂₇, n-C₁₄H₂₉, n-C₁₅H₃₁, n-C₁₆H₃₃, n-C₁₇H₃₅,n-C₁₈H₃₇, n-C₁₉H₃₉ or n-C₂₀H₄₁. In still other embodiments, R₅ is(C₁₀-C₂₀) acyl. In still other embodiments, R₅ is —C(O)n-C₁₀H₂₁,—C(O)n-C₁₁H₂₃, —C(O)n-C₁₀H₂₁, —C(O)n-C₁₂H₂₅, —C(O)n-C₁₃H₂₇,—C(O)n-C₁₄H₂₉, —C(O)n-C₁₅H₃₁, —C(O)n-C₁₆H₃₃, —C(O)n-C₁₇H₃₅,—C(O)n-C₁₈H₃₇, —C(O)n-C₁₉H₃₉ or —C(O)n-C₁₉H₃₉.

A linker (i.e., L), typically contains at least two functional groups,e.g., one functional group that can be used to form a bond with thehydroxyl group of camptothecin or camptothecin derivative and anotherfunctional group that can form a bond with a polymer. Typically, thoughnot necessarily, the functional group on the linker that is used to forma bond with the camptothecin or camptothecin derivative is at one end ofthe linker and the functional group that is used to form a bond with thepolymer is at the other end of the linker.

The linker will typically comprise electrophilic functional groups thatcan react with the nucleophilic hydroxyl group on the camptothecin orcamptothecin derivative to form a covalent bond. The linker can comprisenucleophilic functional groups that can react with electrophilicfunctional groups like carbonyl, halide, or alkoxyl groups on thepolymer. The linker can also comprise electrophilic groups that canreact with nucleophilic functional groups on the polymer. Any of thesebonds may be formed by conventional methods known in the art. Theresulting bonds between the linker, camptothecin or camptothecinderivative and the polymer should be biodegradable.

The linker can be of varying lengths, such as from 1 to 20 atoms inlength. For example, the linker moiety can be from 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 atoms in length,where any of the stated values can form an upper and/or lower end pointof a range. Further, the linker moiety can be substituted orunsubstituted. When substituted, the linker can contain substituentsattached to the backbone of the linker or substituents embedded in thebackbone of the linker. For example, an amine substituted linker moietycan contain an amine group attached to the backbone of the linker or anitrogen in the backbone of the linker.

Suitable linkers include, but are not limited to alkyl, substitutedalkyl, heteroalkyl, substituted heteroalkyl, ethers, esters, polyethers,polyesters, polyalkylenes, polyamines and the like. In a specificexample, the linker can comprise —(CH₂)_(m)—, wherein m is from 1 to 20,and where the point of attachment to the camptothecin or camptothecinderivative is an ether, ester, carbonate or carbamate bond and the pointof attachment to the polymer is an ester, ether, carbamate, amine, oramide bond. For example, the linker moiety can be X¹—(CH₂)_(m)—X²,wherein m is from 1 to 10, and X¹ is —C(O)—, a bond or —C(O)N—, —C(O)O—and X² —C(O), —C(O)O—, —C(O)N—, —NH—, —O— or halo.

In still another embodiment, the linker can comprise a branched orstraight-chain alkyl, wherein one or more of the carbon atoms issubstituted with oxygen (e.g., an ether) or an amino group. For example,suitable linkers can include, but are not limited to, a methoxymethyl,methoxyethyl, methoxypropyl, methoxybutyl, ethoxymethyl, ethoxyethyl,ethoxypropyl, propoxymethyl, propoxyethyl, methylaminomethyl,methylaminoethyl, methylaminopropyl, methylaminobutyl, ethylaminomethyl,ethylaminoethyl, ethylaminopropyl, propylaminomethyl, propylaminoethyl,methoxymethoxymethyl, ethoxymethoxymethyl, methoxyethoxymethyl,methoxymethoxyethyl, and the like, and derivatives thereof, where thepoint of attachment to the hydrophobic drug and/or amino acid is anester, ether, or amide bond.

Exemplary polymers include, but are not limited to poly(D-lactide),poly(L-lactide), poly(D,L-lactide), poly(lactide-co-glycolide),polycaprolactone, poly(lactide-co-caprolactone),poly(glycolide-co-caprolactone), poly(lactic-co-glycolic acid) (PGLA)and/or poly(1,6-bis-p-carboxyphenoxyhexane-co-sebacic anhydride. Thesecopolymers may take the form of random, block, or alternating copolymer,depending on the monomer sequence. Other polymers of use include, butare not limited to, PHMPA, PHMPA-PLMA, PEG, dextran, chitosan andpullan.

Methods for the preparation of compounds of Formulae (I)-(XIV) areconventional and within the ambit of the skilled artisan.

Compositions and Methods of Administration

The compositions provided herein contain therapeutically effectiveamounts of one or more of the compounds provided herein that are usefulin the prevention, treatment, or amelioration of one or more of thesymptoms of diseases or disorders described herein and a vehicle.Vehicles suitable for administration of the compounds provided hereininclude any such carriers known to those skilled in the art to besuitable for the particular mode of administration.

In addition, the compounds may be formulated as the sole activeingredient in the composition or may be combined with other activeingredients.

The compositions contain one or more compounds provided herein. Thecompounds are, in some embodiments, formulated into suitablepreparations such as solutions, suspensions, tablets, dispersibletablets, pills, capsules, powders, sustained release formulations orelixirs, for oral administration or in sterile solutions or suspensionsfor parenteral administration, as well as topical administration,transdermal administration and oral inhalation via nebulizers,pressurized metered dose inhalers and dry powder inhalers. In someembodiments, the compounds described above are formulated intocompositions using techniques and procedures well known in the art (see,e.g., Ansel, Introduction to Pharmaceutical Dosage Forms, SeventhEdition (1999).

In the compositions, effective concentrations of one or more compoundsor derivatives thereof is (are) mixed with a suitable vehicle. Thecompounds may be derivatized as the corresponding salts, esters, enolethers or esters, acetals, ketals, orthoesters, hemiacetals, hemiketals,acids, bases, solvates, ion-pairs, hydrates or prodrugs prior toformulation, as described above. The concentrations of the compounds inthe compositions are effective for delivery of an amount, uponadministration that treats, leads to prevention, or amelioration of oneor more of the symptoms of diseases or disorders described herein. Insome embodiments, the compositions are formulated for single dosageadministration. To formulate a composition, the weight fraction of acompound is dissolved, suspended, dispersed or otherwise mixed in aselected vehicle at an effective concentration such that the treatedcondition is relieved, prevented, or one or more symptoms areameliorated.

The active compound is included in the vehicle in an amount sufficientto exert a therapeutically useful effect in the absence of undesirableside effects on the patient treated. The therapeutically effectiveconcentration may be predicted empirically by testing the compounds inin vitro and in vivo systems well known to those of skill in the art andthen extrapolated therefrom for dosages for humans. Human doses are thentypically fine-tuned in clinical trials and titrated to response.

The concentration of active compound in the composition will depend onabsorption, inactivation and excretion rates of the active compound, thephysicochemical characteristics of the compound, the dosage schedule,and amount administered as well as other factors known to those of skillin the art. For example, the amount that is delivered is sufficient toameliorate one or more of the symptoms of diseases or disorders asdescribed herein.

In some embodiments, a therapeutically effective dosage should produce aserum concentration of active ingredient of from about 0.001 ng/ml toabout 1.0 ng/ml, 2-10 ng/ml, 11 to 50 ng/ml, 51-200 ng/ml, or about 200to 1000 ng/ml. The compositions, in other embodiments, should provide adosage of from about 0.0001 mg to about 70 mg of compound per kilogramof body weight per day. Dosage unit forms are prepared to provide fromabout 0.01 mg, 0.1 mg or 1 mg to about 500 mg, or about 1000 mg, and insome embodiments from about 10 mg to about 500 mg of the activeingredient or a combination of essential ingredients per dosage unitform.

The active ingredient may be administered at once, or may be dividedinto a number of smaller doses to be administered at intervals of time.It is understood that the precise dosage and duration of treatment is afunction of the disease being treated and may be determined empiricallyusing known testing protocols or by extrapolation from in vivo or invitro test data or subsequent clinical testing. It is to be noted thatconcentrations and dosage values may also vary with the severity of thecondition to be alleviated. It is to be further understood that for anyparticular subject, specific dosage regimens should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of thecompositions and that the concentration ranges set forth herein areexemplary only and are not intended to limit the scope or practice ofthe claimed compositions.

In instances in which the compounds exhibit insufficient solubility,methods for solubilizing compounds may be used such as use of liposomes,prodrugs, complexation/chelation, nanoparticles, or emulsions ortertiary templating. Such methods are known to those of skill in thisart, and include, but are not limited to, using co-solvents, such asdimethylsulfoxide (DMSO), using surfactants or surface modifiers, suchas TWEEN®, complexing agents such as cyclodextrin or dissolution byenhanced ionization (i.e. dissolving in aqueous sodium bicarbonate).Derivatives of the compounds, such as prodrugs of the compounds may alsobe used in formulating effective compositions.

Upon mixing or addition of the compound(s), the resulting mixture may bea solution, suspension, emulsion or the like. The form of the resultingmixture depends upon a number of factors, including the intended mode ofadministration and the solubility of the compound in the selectedvehicle. The effective concentration is sufficient for ameliorating thesymptoms of the disease, disorder or condition treated and may beempirically determined.

The compositions are provided for administration to humans and animalsin indication appropriate dosage forms, such as dry powder inhalers(DPIs), pressurized metered dose inhalers (pMDIs), nebulizers, tablets,capsules, pills, sublingual tapes/bioerodible strips, tablets orcapsules, powders, granules, lozenges, lotions, salves, suppositories,fast melts, transdermal patches or other transdermal applicationdevices/preparations, sterile parenteral solutions or suspensions, andoral solutions or suspensions, and oil-water emulsions containingsuitable quantities of the compounds or derivatives thereof. Thetherapeutically active compounds and derivatives thereof are, in someembodiments, formulated and administered in unit-dosage forms ormultiple-dosage forms. Unit-dose forms as used herein refer tophysically discrete units suitable for human and animal subjects andpackaged individually as is known in the art. Each unit-dose contains apredetermined quantity of the therapeutically active compound sufficientto produce the desired therapeutic effect, in association with therequired vehicle. Examples of unit-dose forms include ampoules andsyringes and individually packaged tablets or capsules. Unit-dose formsmay be administered in fractions or multiples thereof. A multiple-doseform is a plurality of identical unit-dosage forms packaged in a singlecontainer to be administered in segregated unit-dose form. Examples ofmultiple-dose forms include vials, bottles of tablets or capsules orbottles of pints or gallons. Hence, multiple dose form is a multiple ofunit-doses which are not segregated in packaging.

Liquid compositions can, for example, be prepared by dissolving,dispersing, or otherwise mixing an active compound as defined above andoptional adjuvants in a vehicle, such as, for example, water, saline,aqueous dextrose, glycerol, glycols, ethanol, and the like, to therebyform a solution or suspension, colloidal dispersion, emulsion orliposomal formulation. If desired, the composition to be administeredmay also contain minor amounts of nontoxic auxiliary substances such aswetting agents, emulsifying agents, solubilizing agents, pH bufferingagents and the like, for example, acetate, sodium citrate, cyclodextrinderivatives, sorbitan monolaurate, triethanolamine sodium acetate,triethanolamine oleate, and other such agents.

Actual methods of preparing such dosage forms are known, or will beapparent, to those skilled in this art; for example, see Remington'sPharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 15thEdition, 1975 or later editions thereof.

Dosage forms or compositions containing active ingredient in the rangeof 0.005% to 100% with the balance made up from vehicle or carrier maybe prepared. Methods for preparation of these compositions are known tothose skilled in the art. The contemplated compositions may contain0.001%-100% active ingredient, in one embodiment 0.1-95%, in anotherembodiment 0.4-10%.

In certain embodiments, the compositions are lactose-free compositionscontaining excipients that are well known in the art and are listed, forexample, in the U.S. Pharmacopeia (USP) 25-NF20 (2002). In general,lactose-free compositions contain active ingredients, a binder/filler,and a lubricant in compatible amounts. Particular lactose-free dosageforms contain active ingredients, microcrystalline cellulose,pre-gelatinized starch, and magnesium stearate.

Further provided are anhydrous compositions and dosage forms comprisingactive ingredients, since water can facilitate the degradation of somecompounds. For example, the addition of water (e.g., 5%) is widelyaccepted as a means of simulating long-term storage in order todetermine characteristics such as shelf-life or the stability offormulations over time. See, e.g., Jens T. Carstensen, Drug Stability:Principles & Practice, 2d. Ed., Marcel Dekker, NY, N.Y., 1995, pp.379-80. In effect, water and heat accelerate the decomposition of somecompounds. Thus, the effect of water on a formulation can be of greatsignificance since moisture and/or humidity are commonly encounteredduring manufacture, handling, packaging, storage, shipment, and use offormulations.

Anhydrous compositions and dosage forms provided herein can be preparedusing anhydrous or low moisture containing ingredients and low moistureor low humidity conditions.

An anhydrous composition should be prepared and stored such that itsanhydrous nature is maintained. Accordingly, anhydrous compositions aregenerally packaged using materials known to prevent exposure to watersuch that they can be included in suitable formulary kits. Examples ofsuitable packaging include, but are not limited to, hermetically sealedfoils, plastics, unit dose containers (e.g., vials), blister packs, andstrip packs.

Oral dosage forms are either solid, gel or liquid. The solid dosageforms are tablets, capsules, granules, and bulk powders. Types of oraltablets include compressed, chewable lozenges and tablets which may beenteric-coated, sugar-coated or film-coated. Capsules may be hard orsoft gelatin capsules, while granules and powders may be provided innon-effervescent or effervescent form with the combination of otheringredients known to those skilled in the art.

In certain embodiments, the formulations are solid dosage forms such asfor example, capsules or tablets. The tablets, pills, capsules, trochesand the like can contain one or more of the following ingredients, orcompounds of a similar nature: a binder; a lubricant; a diluent; aglidant; a disintegrating agent; a coloring agent; a sweetening agent; aflavoring agent; a wetting agent; an enteric coating; a film coatingagent and modified release agent. Examples of binders includemicrocrystalline cellulose, methyl paraben, polyalkyleneoxides, gumtragacanth, glucose solution, acacia mucilage, gelatin solution,molasses, polyvinylpyrrolidine, povidone, crospovidones, sucrose andstarch and starch derivatives. Lubricants include talc, starch,magnesium/calcium stearate, lycopodium and stearic acid. Diluentsinclude, for example, lactose, sucrose, trehalose, lysine, leucine,lecithin, starch, kaolin, salt, mannitol and dicalcium phosphate.Glidants include, but are not limited to, colloidal silicon dioxide.Disintegrating agents include crosscarmellose sodium, sodium starchglycolate, alginic acid, corn starch, potato starch, bentonite,methylcellulose, agar and carboxymethylcellulose. Coloring agentsinclude, for example, any of the approved certified water soluble FD andC dyes, mixtures thereof; and water insoluble FD and C dyes suspended onalumina hydrate and advanced coloring or anti-forgery color/opalescentadditives known to those skilled in the art. Sweetening agents includesucrose, lactose, mannitol and artificial sweetening agents such assaccharin, and any number of spray dried flavors. Flavoring agentsinclude natural flavors extracted from plants such as fruits andsynthetic blends of compounds which produce a pleasant sensation or maskunpleasant taste, such as, but not limited to peppermint and methylsalicylate. Wetting agents include propylene glycol monostearate,sorbitan monooleate, diethylene glycol monolaurate and polyoxyethylenelauryl ether. Enteric-coatings include fatty acids, fats, waxes,shellac, ammoniated shellac and cellulose acetate phthalates. Filmcoatings include hydroxyethylcellulose, sodium carboxymethylcellulose,polyethylene glycol 4000 and cellulose acetate phthalate. Modifiedrelease agents include polymers such as the Eudragit® series andcellulose esters.

The compound, or derivative thereof, can be provided in a compositionthat protects it from the acidic environment of the stomach. Forexample, the composition can be formulated in an enteric coating thatmaintains its integrity in the stomach and releases the active compoundin the intestine. The composition may also be formulated in combinationwith an antacid or other such ingredient.

When the dosage unit form is a capsule, it can contain, in addition tomaterial of the above type, a liquid carrier such as a fatty oil. Inaddition, dosage unit forms can contain various other materials whichmodify the physical form of the dosage unit, for example, coatings ofsugar and other enteric agents. The compounds can also be administeredas a component of an elixir, suspension, syrup, wafer, sprinkle, chewinggum or the like. A syrup may contain, in addition to the activecompounds, sucrose as a sweetening agent and certain preservatives, dyesand colorings and flavors.

The active materials can also be mixed with other active materials whichdo not impair the desired action, or with materials that supplement thedesired action, such as antacids, H₂ blockers, and diuretics. The activeingredient is a compound or derivative thereof as described herein.Higher concentrations, up to about 98% by weight of the activeingredient may be included.

In all embodiments, tablets and capsules formulations may be coated asknown by those of skill in the art in order to modify or sustaindissolution of the active ingredient. Thus, for example, they may becoated with a conventional enterically digestible coating, such asphenylsalicylate, waxes and cellulose acetate phthalate.

Liquid oral dosage forms include aqueous solutions, emulsions,suspensions, solutions and/or suspensions reconstituted fromnon-effervescent granules and effervescent preparations reconstitutedfrom effervescent granules. Aqueous solutions include, for example,elixirs and syrups. Emulsions are either oil-in-water or water-in-oil.

Elixirs are clear, sweetened, hydroalcoholic preparations. Vehicles usedin elixirs include solvents. Syrups are concentrated aqueous solutionsof a sugar, for example, sucrose, and may contain a preservative. Anemulsion is a two-phase system in which one liquid is dispersed in theform of small globules throughout another liquid. Carriers used inemulsions are non-aqueous liquids, emulsifying agents and preservatives.Suspensions use suspending agents and preservatives. Acceptablesubstances used in non-effervescent granules, to be reconstituted into aliquid oral dosage form, include diluents, sweeteners and wettingagents. Acceptable substances used in effervescent granules, to bereconstituted into a liquid oral dosage form, include organic acids anda source of carbon dioxide. Coloring and flavoring agents are used inall of the above dosage forms.

Solvents include glycerin, sorbitol, ethyl alcohol and syrup. Examplesof preservatives include glycerin, methyl and propylparaben, benzoicacid, sodium benzoate and alcohol. Examples of non-aqueous liquidsutilized in emulsions include mineral oil and cottonseed oil. Examplesof emulsifying agents include gelatin, acacia, tragacanth, bentonite,and surfactants such as polyoxyethylene sorbitan monooleate. Suspendingagents include sodium carboxymethylcellulose, pectin, tragacanth, Veegumand acacia. Sweetening agents include sucrose, syrups, glycerin andartificial sweetening agents such as saccharin. Wetting agents includepropylene glycol monostearate, sorbitan monooleate, diethylene glycolmonolaurate and polyoxyethylene lauryl ether. Organic acids includecitric and tartaric acid. Sources of carbon dioxide include sodiumbicarbonate and sodium carbonate. Coloring agents include any of theapproved certified water soluble FD and C dyes, and mixtures thereof.Flavoring agents include natural flavors extracted from plants suchfruits, and synthetic blends of compounds which produce a pleasant tastesensation.

For a solid dosage form, the solution or suspension, in for example,propylene carbonate, vegetable oils or triglycerides, is in someembodiments encapsulated in a gelatin capsule. Such solutions, and thepreparation and encapsulation thereof, are disclosed in U.S. Pat. Nos.4,328,245; 4,409,239; and 4,410,545. For a liquid dosage form, thesolution, e.g., for example, in a polyethylene glycol, may be dilutedwith a sufficient quantity of a liquid vehicle, e.g., water, to beeasily measured for administration.

Alternatively, liquid or semi-solid oral formulations may be prepared bydissolving or dispersing the active compound or salt in vegetable oils,glycols, triglycerides, propylene glycol esters (e.g., propylenecarbonate) and other such carriers, and encapsulating these solutions orsuspensions in hard or soft gelatin capsule shells. Other usefulformulations include those set forth in U.S. Pat. Nos. RE 28,819 and4,358,603. Briefly, such formulations include, but are not limited to,those containing a compound provided herein, a dialkylated mono- orpolyalkylene glycol, including, but not limited to, 1,2-dimethoxyethane,diglyme, triglyme, tetraglyme, polyethylene glycol-350-dimethyl ether,polyethylene glycol-550-dimethyl ether, polyethylene glycol-750-dimethylether wherein 350, 550 and 750 refer to the approximate averagemolecular weight of the polyethylene glycol, and one or moreantioxidants, such as butylated hydroxytoluene (BHT), butylatedhydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone,hydroxycoumarins, ethanolamine, lecithin, cephalin, ascorbic acid, malicacid, sorbitol, phosphoric acid, thiodipropionic acid and its esters,and dithiocarbamates.

Other formulations include, but are not limited to, aqueous alcoholicsolutions including an acetal. Alcohols used in these formulations areany water-miscible solvents having one or more hydroxyl groups,including, but not limited to, propylene glycol and ethanol. Acetalsinclude, but are not limited to, di(lower alkyl) acetals of lower alkylaldehydes such as acetaldehyde diethyl acetal.

Parenteral administration, in some embodiments characterized byinjection, either subcutaneously, intramuscularly or intravenously isalso contemplated herein. Injectables can be prepared in conventionalforms, either as liquid solutions or suspensions, solid forms suitablefor solution or suspension in liquid prior to injection, or asemulsions. The injectables, solutions and emulsions also contain one ormore excipients. Suitable excipients are, for example, water, saline,dextrose, glycerol or ethanol. In addition, if desired, the compositionsto be administered may also contain minor amounts of non-toxic auxiliarysubstances such as wetting or emulsifying agents, pH buffering agents,stabilizers, solubility enhancers, and other such agents, such as forexample, sodium acetate, sorbitan monolaurate, triethanolamine oleateand cyclodextrins.

Implantation of a slow-release or sustained-release system, such that aconstant level of dosage is maintained (see, e.g., U.S. Pat. No.3,710,795) is also contemplated herein. Briefly, a compound providedherein is dispersed in a solid inner matrix, e.g.,polymethylmethacrylate, polybutylmethacrylate, plasticized orunplasticized polyvinylchloride, plasticized nylon, plasticizedpolyethyleneterephthalate, natural rubber, polyisoprene,polyisobutylene, polybutadiene, polyethylene, ethylene-vinylacetatecopolymers, silicone rubbers, polydimethylsiloxanes, silicone carbonatecopolymers, hydrophilic polymers such as hydrogels of esters of acrylicand methacrylic acid, collagen, cross-linked polyvinylalcohol andcross-linked partially hydrolyzed polyvinyl acetate, that is surroundedby an outer polymeric membrane, e.g., polyethylene, polypropylene,ethylene/propylene copolymers, ethylene/ethyl acrylate copolymers,ethylene/vinylacetate copolymers, silicone rubbers, polydimethylsiloxanes, neoprene rubber, chlorinated polyethylene, polyvinylchloride,vinylchloride copolymers with vinyl acetate, vinylidene chloride,ethylene and propylene, ionomer polyethylene terephthalate, butyl rubberepichlorohydrin rubbers, ethylene/vinyl alcohol copolymer,ethylene/vinyl acetate/vinyl alcohol terpolymer, andethylene/vinyloxyethanol copolymer, that is insoluble in body fluids.The compound diffuses through the outer polymeric membrane in a releaserate controlling step. The percentage of active compound contained insuch parenteral compositions is highly dependent on the specific naturethereof, as well as the activity of the compound and the needs of thesubject.

Parenteral administration of the compositions includes intravenous,subcutaneous and intramuscular administrations. Preparations forparenteral administration include sterile solutions ready for injection,sterile dry soluble products, such as lyophilized powders, ready to becombined with a solvent just prior to use, including hypodermic tablets,sterile suspensions ready for injection, sterile dry insoluble productsready to be combined with a vehicle just prior to use and sterileemulsions. The solutions may be either aqueous or nonaqueous.

If administered intravenously, suitable carriers include physiologicalsaline or phosphate buffered saline (PBS), and solutions containingthickening and solubilizing agents, such as glucose, polyethyleneglycol, and polypropylene glycol and mixtures thereof.

Vehicles used in parenteral preparations include aqueous vehicles,nonaqueous vehicles, antimicrobial agents, isotonic agents, buffers,antioxidants, local anesthetics, suspending and dispersing agents,emulsifying agents, sequestering or chelating agents and othersubstances.

Examples of aqueous vehicles include Sodium Chloride Injection, RingersInjection, Isotonic Dextrose Injection, Sterile Water Injection,Dextrose and Lactated Ringers Injection. Nonaqueous parenteral vehiclesinclude fixed oils of vegetable origin, cottonseed oil, corn oil, sesameoil and peanut oil. Antimicrobial agents in bacteriostatic orfungistatic concentrations must be added to parenteral preparationspackaged in multiple-dose containers which include phenols or cresols,mercurials, benzyl alcohol, chlorobutanol, methyl and propylp-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride andbenzethonium chloride. Isotonic agents include sodium chloride anddextrose. Buffers include phosphate and citrate. Antioxidants includesodium bisulfate. Local anesthetics include procaine hydrochloride.Suspending and dispersing agents include sodium carboxymethylcellulose,hydroxypropyl methylcellulose and polyvinylpyrrolidone. Emulsifyingagents include Polysorbate 80 (Tween® 80). A sequestering or chelatingagent of metal ions includes EDTA. Carriers also include ethyl alcohol,polyethylene glycol and propylene glycol for water miscible vehicles;and sodium hydroxide, hydrochloric acid, citric acid or lactic acid forpH adjustment.

The concentration of compound is adjusted so that an injection providesan effective amount to produce the desired pharmacological effect. Theexact dose depends on the age, weight, body surface area and conditionof the patient or animal as is known in the art.

The unit-dose parenteral preparations are packaged in an ampoule, a vialor a syringe with a needle. All preparations for parenteraladministration must be sterile, as is known and practiced in the art.

Illustratively, intravenous or intraarterial infusion of a sterileaqueous solution containing an active compound is an effective mode ofadministration. Another embodiment is a sterile aqueous or oily solutionor suspension containing an active material injected as necessary toproduce the desired pharmacological effect.

Injectables are designed for local and systemic administration. In someembodiments, a therapeutically effective dosage is formulated to containa concentration of at least about 0.01% w/w up to about 90% w/w or more,in certain embodiments more than 0.1% w/w of the active compound to thetreated tissue(s).

The compound may be suspended in micronized or other suitable form ormay be derivatized to produce a more soluble active product or toproduce a prodrug. The form of the resulting mixture depends upon anumber of factors, including the intended mode of administration and thesolubility of the compound in the selected carrier or vehicle. Theeffective concentration is sufficient for ameliorating the symptoms ofthe condition and may be empirically determined.

Active ingredients provided herein can be administered by controlledrelease means or by delivery devices that are well known to those ofordinary skill in the art. Examples include, but are not limited to,those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809;3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548;5,073,543; 5,639,476; 5,354,556; 5,639,480; 5,733,566; 5,739,108;5,891,474; 5,922,356; 5,972,891; 5,980,945; 5,993,855; 6,045,830;6,087,324; 6,113,943; 6,197,350; 6,248,363; 6,264,970; 6,267,981;6,376,461; 6,419,961; 6,589,548; 6,613,358; 6,699,500 and 6,740,634.Such dosage forms can be used to provide slow or controlled-release ofone or more active ingredients using, for example, hydroxypropylmethylcellulose, other polymer matrices, gels, permeable membranes, osmoticsystems, multilayer coatings, microparticles, liposomes, microspheres,or a combination thereof to provide the desired release profile invarying proportions. Suitable controlled-release formulations known tothose of ordinary skill in the art, including those described herein,can be readily selected for use with the active ingredients providedherein.

All controlled-release products have a common goal of improving drugtherapy over that achieved by their non-controlled counterparts.Ideally, the use of an optimally designed controlled-release preparationin medical treatment is characterized by a minimum of drug substancebeing employed to cure or control the condition in a minimum amount oftime. Advantages of controlled-release formulations include extendedactivity of the drug, reduced dosage frequency, and increased patientcompliance. In addition, controlled-release formulations can be used toaffect the time of onset of action or other characteristics, such asblood levels of the drug, and can thus affect the occurrence of side(e.g., adverse) effects.

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled-release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or compounds.

In certain embodiments, the agent may be administered using intravenousinfusion, an implantable osmotic pump, a transdermal patch, liposomes,or other modes of administration. In some embodiments, a pump may beused (see, Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwaldet al., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574(1989)). In other embodiments, polymeric materials can be used. In otherembodiments, a controlled release system can be placed in proximity ofthe therapeutic target, i.e., thus requiring only a fraction of thesystemic dose (see, e.g., Goodson, Medical Applications of ControlledRelease, vol. 2, pp. 115-138 (1984)). In some embodiments, a controlledrelease device is introduced into a subject in proximity of the site ofinappropriate immune activation or a tumor. Other controlled releasesystems are discussed in the review by Langer (Science 249:1527-1533(1990)). The active ingredient can be dispersed in a solid inner matrix,e.g., polymethylmethacrylate, polybutylmethacrylate, plasticized orunplasticized polyvinylchloride, plasticized nylon, plasticizedpolyethyleneterephthalate, natural rubber, polyisoprene,polyisobutylene, polybutadiene, polyethylene, ethylene-vinylacetatecopolymers, silicone rubbers, polydimethylsiloxanes, silicone carbonatecopolymers, hydrophilic polymers such as hydrogels of esters of acrylicand methacrylic acid, collagen, cross-linked polyvinylalcohol andcross-linked partially hydrolyzed polyvinyl acetate, that is surroundedby an outer polymeric membrane, e.g., polyethylene, polypropylene,ethylene/propylene copolymers, ethylene/ethyl acrylate copolymers,ethylene/vinylacetate copolymers, silicone rubbers, polydimethylsiloxanes, neoprene rubber, chlorinated polyethylene, polyvinylchloride,vinylchloride copolymers with vinyl acetate, vinylidene chloride,ethylene and propylene, ionomer polyethylene terephthalate, butyl rubberepichlorohydrin rubbers, ethylene/vinyl alcohol copolymer,ethylene/vinyl acetate/vinyl alcohol terpolymer, andethylene/vinyloxyethanol copolymer, that is insoluble in body fluids.The active ingredient then diffuses through the outer polymeric membranein a release rate controlling step. The percentage of active ingredientcontained in such parenteral compositions is highly dependent on thespecific nature thereof, as well as the needs of the subject.

Of interest, herein are also lyophilized powders, which can bereconstituted for administration as solutions, emulsions and othermixtures. They may also be reconstituted and formulated as solids orgels.

The sterile, lyophilized powder is prepared by dissolving a compoundprovided herein, or a derivative thereof, in a suitable solvent. Thesolvent may contain an excipient which improves the stability or otherpharmacological component of the powder or reconstituted solution,prepared from the powder. Excipients that may be used include, but arenot limited to, an antioxidant, a buffer and a bulking agent. In someembodiments, the excipient is selected from dextrose, sorbitol,fructose, corn syrup, xylitol, glycerin, glucose, sucrose and othersuitable agents. The solvent may contain a buffer, such as citrate,sodium or potassium phosphate or other such buffer known to those ofskill in the art at, at about neutral pH. Subsequent sterile filtrationof the solution followed by lyophilization under standard conditionsknown to those of skill in the art provides the desired formulation. Insome embodiments, the resulting solution will be apportioned into vialsfor lyophilization. Each vial will contain a single dosage or multipledosages of the compound. The lyophilized powder can be stored underappropriate conditions, such as at about 4° C. to room temperature.

Reconstitution of this lyophilized powder with water for injectionprovides a formulation for use in parenteral administration. Forreconstitution, the lyophilized powder is added to sterile water orother suitable carriers. The precise amount depends upon the selectedcompound. Such amount can be empirically determined.

Topical mixtures are prepared as described for the local and systemicadministration. The resulting mixture may be a solution, suspension,emulsions or the like and are formulated as creams, gels, ointments,emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes,foams, aerosols, irrigations, sprays, suppositories, bandages, dermalpatches or any other formulations suitable for topical administration.

The compounds or derivatives thereof may be formulated as aerosols fortopical application, such as by inhalation (see, e.g., U.S. Pat. Nos.4,044,126, 4,414,209, and 4,364,923, which describe aerosols fordelivery of a steroid useful for treatment of inflammatory diseases,particularly asthma). These formulations for administration to therespiratory tract can be in the form of an aerosol or solution for anebulizer, or as a microfine powder for insufflation, alone or incombination with an inert carrier such as lactose. In such a case, theparticles of the formulation will, in some embodiments, have mass mediangeometric diameters of less than 5 microns, in other embodiments lessthan 10 microns.

Oral inhalation formulations of the compounds or derivatives suitablefor inhalation include metered dose inhalers, dry powder inhalers andliquid preparations for administration from a nebulizer or metered doseliquid dispensing system. For both metered dose inhalers and dry powderinhalers, a crystalline form of the compounds or derivatives is thepreferred physical form of the drug to confer longer product stability.

In addition to particle size reduction methods known to those skilled inthe art, crystalline particles of the compounds or derivatives can begenerated using supercritical fluid processing which offers significantadvantages in the production of such particles for inhalation deliveryby producing respirable particles of the desired size in a single step.(e.g., International Publication No. WO2005/025506). A controlledparticle size for the microcrystals can be selected to ensure that asignificant fraction of the compounds or derivatives is deposited in thelung. In some embodiments, these particles have a mass medianaerodynamic diameter of about 0.1 to about 10 microns, in otherembodiments, about 1 to about 5 microns and still other embodiments,about 1.2 to about 3 microns.

Inert and non-flammable HFA propellants are selected from HFA 134a(1,1,1,2-tetrafluoroethane) and HFA 227e(1,1,1,2,3,3,3-heptafluoropropane) and provided either alone or as aratio to match the density of crystal particles of the compounds orderivatives. A ratio is also selected to ensure that the productsuspension avoids detrimental sedimentation or cream (which canprecipitate irreversible agglomeration) and instead promote a looselyflocculated system, which is easily dispersed when shaken. Looselyfluctuated systems are well regarded to provide optimal stability forpMDI canisters. As a result of the formulation's properties, theformulation contained no ethanol and no surfactants/stabilizing agents.

The compounds may be formulated for local or topical application, suchas for topical application to the skin and mucous membranes, such as inthe eye, in the form of gels, creams, and lotions and for application tothe eye or for intracisternal or intraspinal application. Topicaladministration is contemplated for transdermal delivery and also foradministration to the eyes or mucosa, or for inhalation therapies. Nasalsolutions of the active compound alone or in combination with otherexcipients can also be administered.

For nasal administration, the preparation may contain an esterifiedphosphonate compound dissolved or suspended in a liquid carrier, inparticular, an aqueous carrier, for aerosol application. The carrier maycontain solubilizing or suspending agents such as propylene glycol,surfactants, absorption enhancers such as lecithin or cyclodextrin, orpreservatives.

Solutions, particularly those intended for ophthalmic use, may beformulated as 0.01%-10% isotonic solutions, pH about 5-7.4, withappropriate salts.

Other routes of administration, such as transdermal patches, includingiontophoretic and electrophoretic devices, and rectal administration,are also contemplated herein.

Transdermal patches, including iontophoretic and electrophoreticdevices, are well known to those of skill in the art. For example, suchpatches are disclosed in U.S. Pat. Nos. 6,267,983, 6,261,595, 6,256,533,6,167,301, 6,024,975, 6,010,715, 5,985,317, 5,983,134, 5,948,433 and5,860,957.

For example, dosage forms for rectal administration are rectalsuppositories, capsules and tablets for systemic effect. Rectalsuppositories are used herein mean solid bodies for insertion into therectum which melt or soften at body temperature releasing one or morepharmacologically or therapeutically active ingredients. Substancesutilized in rectal suppositories are bases or vehicles and agents toraise the melting point. Examples of bases include cocoa butter(theobroma oil), glycerin-gelatin, carbowax (polyoxyethylene glycol) andappropriate mixtures of mono-, di- and triglycerides of fatty acids.Combinations of the various bases may be used. Agents to raise themelting point of suppositories include spermaceti and wax. Rectalsuppositories may be prepared either by the compressed method or bymolding. The weight of a rectal suppository, in one embodiment, is about2 to 3 gm. Tablets and capsules for rectal administration aremanufactured using the same substance and by the same methods as forformulations for oral administration.

The compounds provided herein, or derivatives thereof, may also beformulated to be targeted to a particular tissue, receptor, or otherarea of the body of the subject to be treated. Many such targetingmethods are well known to those of skill in the art. All such targetingmethods are contemplated herein for use in the instant compositions. Fornon-limiting examples of targeting methods, see, e.g., U.S. Pat. Nos.6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570,6,120,751, 6,071,495, 6,060,082, 6,048,736, 6,039,975, 6,004,534,5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542 and 5,709,874.

In some embodiments, liposomal suspensions, including tissue-targetedliposomes, such as tumor-targeted liposomes, may also be suitable ascarriers. These may be prepared according to methods known to thoseskilled in the art. For example, liposome formulations may be preparedas described in U.S. Pat. No. 4,522,811. Briefly, liposomes such asmultilamellar vesicles (MLV's) may be formed by drying down phosphatidylcholine and phosphatidyl serine (7:3 molar ratio) on the inside of aflask. A solution of a compound provided herein in phosphate bufferedsaline lacking divalent cations (PBS) is added and the flask shakenuntil the lipid film is dispersed. The resulting vesicles are washed toremove unencapsulated compound, pelleted by centrifugation, and thenresuspended in PBS.

The compounds described herein may be formulated in compositionsdescribed in the art for delivery of hydrophobic medicines (see, e.g.,U.S. Pat. Nos. 5,645,856, 6,096,338, 5,510,103, 5,955,509, 6,322,817,6,555,139, 6,579,519, 6,746,653, 6,834,791, 7,094,810, 8,715,741 and8,414,926) or as composite nanoparticles described in U.S. ProvisionalPatent No. 62/305,526.

The compounds or derivatives may be packaged as articles of manufacturecontaining packaging material, a compound or derivative thereof providedherein, which is effective for treatment, prevention or amelioration ofone or more symptoms of the diseases or disorders, supra, within thepackaging material, and a label that indicates that the compound orcomposition or derivative thereof, is used for the treatment, preventionor amelioration of one or more symptoms of the diseases or disorders,supra.

The articles of manufacture provided herein contain packaging materials.Packaging materials for use in packaging products are well known tothose of skill in the art. See, e.g., U.S. Pat. Nos. 5,323,907,5,052,558 and 5,033,252. Examples of packaging materials include, butare not limited to, blister packs, bottles, tubes, inhalers, pumps,bags, vials, containers, syringes, bottles, and any packaging materialsuitable for a selected formulation and intended mode of administrationand treatment. A wide array of formulations of the compounds andcompositions provided herein are contemplated as are a variety oftreatments for any disease or disorder described herein.

Dosages

In human therapeutics, the physician will determine the dosage regimenthat is most appropriate according to a preventive or curative treatmentand according to the age, weight, stage of the disease and other factorsspecific to the subject to be treated. The compositions, in otherembodiments, should provide a dosage of from about 0.0001 mg to about 70mg of compound per kilogram of body weight per day. Dosage unit formsare prepared to provide from about 0.01 mg, 0.1 mg or 1 mg to about 500mg, or about 1000 mg, and in some embodiments from about 10 mg to about500 mg of the active ingredient or a combination of essentialingredients per dosage unit form.

The amount of active ingredient in the formulations provided herein,which will be effective in the prevention or treatment of a disorder orone or more symptoms thereof, will vary with the nature and severity ofthe disease or condition, and the route by which the active ingredientis administered. The frequency and dosage will also vary according tofactors specific for each subject depending on the specific therapy(e.g., therapeutic or prophylactic agents) administered, the severity ofthe disorder, disease, or condition, the route of administration, aswell as age, body, weight, response, and the past medical history of thesubject.

Exemplary doses of a formulation include milligram or microgram amountsof the active compound per kilogram of subject (e.g., from about 1micrograms per kilogram to about 50 milligrams per kilogram, from about10 micrograms per kilogram to about 30 milligrams per kilogram, fromabout 100 micrograms per kilogram to about 10 milligrams per kilogram,or from about 100 microgram per kilogram to about 5 milligrams perkilogram).

It may be necessary to use dosages of the active ingredient outside theranges disclosed herein in some cases, as will be apparent to those ofordinary skill in the art. Furthermore, it is noted that the clinicianor treating physician will know how and when to interrupt, adjust, orterminate therapy in conjunction with subject response.

Different therapeutically effective amounts may be applicable fordifferent diseases and conditions, as will be readily known by those ofordinary skill in the art. Similarly, amounts sufficient to prevent,manage, treat or ameliorate such disorders, but insufficient to cause,or sufficient to reduce, adverse effects associated with the compositionprovided herein are also encompassed by the above described dosageamounts and dose frequency schedules. Further, when a subject isadministered multiple dosages of a composition provided herein, not allof the dosages need be the same. For example, the dosage administered tothe subject may be increased to improve the prophylactic or therapeuticeffect of the composition or it may be decreased to reduce one or moreside effects that a particular subject is experiencing.

In certain embodiments, administration of the same formulation providedherein may be repeated and the administrations may be separated by atleast 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days,2 months, 75 days, 3 months, or 6 months.

Methods of Use of the Compounds and Compositions

Methods of treating, preventing, or ameliorating one or more symptoms ofdiseases including, for example, cancer and infection also providedherein. The compounds described herein may be used to treat or preventcancers and/or tumors of the anus, bile duct, bladder, bone, bonemarrow, bowel (including colon and rectum), breast, eye, gall bladder,kidney, mouth, larynx, esophagus, stomach, testis, cervix, head, neck,ovary, lung, mesothelioma, neuroendocrine, penis, skin, spinal cord,thyroid, vagina, vulva, uterus, liver, muscle, pancreas, prostate, bloodcells (including lymphocytes and other immune system cells), and brain.Specific cancers contemplated for treatment include carcinomas,Karposi's sarcoma, melanoma, mesothelioma, soft tissue sarcoma,pancreatic cancer, lung cancer, leukemia (acute lymphoblastic, acutemyeloid, chronic lymphocytic, chronic myeloid, and other), and lymphoma(Hodgkin's and non-Hodgkin's), and multiple myeloma.

Other examples of cancers that can be treated according to the methodsdisclosed herein are adrenocortical carcinoma, adrenocortical carcinoma,cerebellar astrocytoma, basal cell carcinoma, bile duct cancer, bladdercancer, bone cancer, brain tumor, breast cancer, Burkitt's lymphoma,carcinoid tumor, central nervous system lymphoma, cervical cancer,chronic myeloproliferative disorders, colon cancer, cutaneous T-celllymphoma, endometrial cancer, ependymoma, esophageal cancer, gallbladdercancer, gastric (stomach) cancer, gastrointestinal carcinoid tumor, germcell tumor, glioma, hairy cell leukemia, head and neck cancer,hepatocellular (liver) cancer, hypopharyngeal cancer, hypothalamic andvisual pathway glioma, intraocular melanoma, retinoblastoma, islet cellcarcinoma (endocrine pancreas), laryngeal cancer, lip and oral cavitycancer, liver cancer, medulloblastoma, Merkel cell carcinoma, squamousneck cancer with occult mycosis fungoides, myelodysplastic syndromes,myelogenous leukemia, nasal cavity and paranasal sinus cancer,nasopharyngeal cancer, neuroblastoma, non-small cell lung cancer, oralcancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreaticcancer, paranasal sinus and nasal cavity cancer, parathyroid cancer,penile cancer, pheochromocytoma, pineoblastoma and supratentorialprimitive neuroectodermal tumor, pituitary tumor, plasma cellneoplasm/multiple myeloma, pleuropulmonary blastoma, prostate cancer,rectal cancer, renal cell (kidney) cancer, retinoblastoma,rhabdomyosarcoma, salivary gland cancer, Ewing's sarcoma, soft tissuesarcoma, Sezary syndrome, skin cancer, small cell lung cancer, smallintestine cancer, supratentorial primitive neuroectodermal tumors,testicular cancer, thymic carcinoma, thymoma, thyroid cancer,transitional cell cancer of the renal pelvis and ureter, trophoblastictumor, urethral cancer, uterine cancer, vaginal cancer, vulvar cancer,Waldenström's macroglobulinemia, and Wilms' tumor.

In practicing the methods, therapeutically effective amounts of thecompounds or compositions, described herein, supra, are administered.

Combination Therapy

The compounds and compositions disclosed herein may also be used incombination with one or more other active ingredients. In certainembodiments, the compounds may be administered in combination, orsequentially, with another therapeutic agent. Such other therapeuticagents include those known for treatment, prevention, or amelioration ofone or more symptoms associated with, for example cancer.

It should be understood that any suitable combination of the compoundsand compositions provided herein with one or more of the abovetherapeutic agents and optionally one or more further pharmacologicallyactive substances are considered to be within the scope of the presentdisclosure. In some embodiments, the compounds and compositions providedherein are administered prior to or subsequent to the one or moreadditional active ingredients.

Finally, it should be noted that there are alternative ways ofimplementing the present invention. Accordingly, the present embodimentsare to be considered as illustrative and not restrictive, and theinvention is not to be limited to the details given herein, but may bemodified within the scope and equivalents of the appended claims.

All publications and patents cited herein are incorporated by referencein their entirety.

The following examples are provided for illustrative purposes only andare not intended to limit the scope of the invention.

EXAMPLES Example 1 Preparation of Compound 1

To a solution of SN38 (1 g, 2.55 mmol, 1.0 eq.) in dichloromethane (400mL) were added pyridine (5 mL) and Boc anhydride (6.39 g, 6.371 mmol,2.5 eq.) and the reaction mixture was stirred at room temperature for 36h. The reaction mixture was filtered and washed with 0.01N HCl (3×). Theorganic phase was separated, dried (sodium sulfate) and evaporated underreduced pressure to provide compound 1 (1.26 g), which was used in thenext step without further purification.

Example 2 Preparation of Compound 2

To a solution of compound 1 (630 mg, 1.3 mmol, 1.0 eq) indichloremethane (20 mL) were added N,N′-dicyclohexylcarbodiimide (DCC)(538 mg, 2.60 mmol, 2.0 eq), DMAP (190 mg, 1.56 mmol, 1.2 eq) andvaleric acid (265 mg, 2.6 mmol 2.0 eq) and the reaction mixture wasstirred at room temperature for 24 h.

The mixture was then filtered, dried (MgSO4) and solvent was evaporatedto provide a residue, which was purified by column chromatography toyield the valerate ester of compound 1. The ester was then treated with80% TFA in DCM and stirred at room temperature for 3 h. TFA was removedand the crude was purified by column chromatography to provide compound2 (LGP001).

Example 3 Preparation of Compound 3

To a solution of compound 1 (630 mg, 1.3 mmol, 1.0 eq) indichloromethane (20 mL) were added N,N′-dicyclohexylcarbodiimide (DCC)(538 mg, 2.60 mmol, 2.0 eq), DMAP (190 mg, 1.56 mmol, 1.2 eq) andtridecanoic acid (556 mg, 2.6 mmol 2.0 eq) and the reaction mix wasstirred at room temperature for 24 h.

The mixture was then filtered, dried (MgSO4) and solvent was evaporatedto provide a residue, which was purified by column chromatography toyield BOC-SN38 ester. The ester was then treated with 80% TFA in DCM andstirred at room temperature for 3 h. TFA was removed and the crude waspurified by column chromatography to provide compound 3 (LGP002).

Example 4 Preparation of Compound 4

To a solution of PLGA-NHS (100 mg, 0.0066 mmol, 1.0 eq, Akina, Inc.,3495, Kent Ave. West Lafayette, Ind. 47906, cat.# A1096, approximatemolecular weight of 15000) in DMF (4 mL) was added hydroxyl compound 2(4.71 mg, 0.0099 mmol, 1.5 eq.) and DIEA (0.1 mL). The reaction mixturewas stirred at room temperature for 12 h, diluted with ether toprecipitate the polymer, placed in the freezer overnight and thesolvents were decanted to provide a yellow viscous material. The yellowmaterial was washed with ether (2×) and the flask was placed undervacuum to provide a yellow foamy material.

Example 5 Anti-Proliferative Activity of Compounds 2 and 3 AgainstVarious Cell Lines

Human colorectal adenocarcinoma cell lines HT29 and SW620, as well asmurine colon carcinoma cell line CT26, were purchased from the AmericanType Culture Collection (ATCC) and grown in either Dulbecco's ModifiedEagle Medium (DMEM) or Roswell Park Memorial Institute (RPMI) mediumfrom Corning and Life Technologies, respectively. These media weresupplemented with 10% fetal bovine serum (FBS, Atlanta Biologicals), aswell as 1 mM sodium pyruvate (Corning) and 10 mM HEPES (Fisher) for CT26cells. Cells were grown in Corning 75 cm² flasks and split approximately6 days before plating, with a refeed of fresh media the day prior toplating. Anti-proliferative drugs 2, 3 and SN38, were reconstituted inDMSO for storage at −20° C. until use. AlamarBlue, a colorimetric cellmetabolism reagent, was purchased from Life Technologies.

To qualify AlamarBlue incubation time and required cell density, cellswere plated into flat bottom 96 well plates (Techno Plastic Products,Trasadingen, Switzerland) at plating densities from 4×10⁴ to 1.25×10³cells per well. The cells were incubated at 37° C. inside a Nu-Aire TSAutoflow incubator with 5% CO₂ and humidity for 48-72 hours, AlamarBluereagent was then added to all wells, equivalent to 10% of the culturevolume or 20 μL. The plates were returned to the 37° C. incubator, andthe absorbance at 570 and 600 nm were assessed using a Bio-Tek Epochmicroplate reader at 1, 2, 4, 7 and in some experiment 24 hours postaddition of AlamarBlue. Once the optimum cell density and incubationtime was determined for each cell line, testing of theanti-proliferative drugs on the cells was initiated. For each cell line,cells were plated at 5×10³ cells per well and incubated at 37° C. for 24hours, at which time the anti-proliferative compounds were added.Preliminary studies showed an optimum dose response of inhibition with 2and 2 using doses from 100 μM to 0.4 μM. Aliquots of anti-proliferativeswere thawed, diluted in media appropriate for each cell line, andserially diluted 1:3. A serial pipetter, was used to deliver 100 μl ofeach concentration of anti-proliferative compound to the appropriatewells in triplicate, the plate was tapped gently to mix, and thenincubated at 37° C. for 48-72 hours. Control wells containing mediaalone and media with an equal percentage of DMSO in comparison to theinitial anti-proliferative dilution were also established in each plate.After a minimum of 48 hours of incubation, 20 μl of AlamarBlue was addedto each experimental well as well as the negative control wells.AlamarBlue positive control wells, containing 200 μl of 100% reducedAlamarBlue, were also established to provide a colorimetric endpointreference for the assay. After addition of AlamarBlue, plates wereincubated for 4 hours at 37° C. and absorbance assessed using the Epochmicroplate reader. Absorbances were used to calculate the % inhibitionfor each well using the following equation, where O1 is the molarextinction coefficient of oxidized AlamarBlue at 570 nm (80586), O2 isthe molar extinction coefficient of oxidized AlamarBlue at 600 nm(117216), A1 is the absorbance of the test wells at 570 nm, A2 is theabsorbance of the test wells at 600 nm, P1 is the absorbance of thecontrol wells with AlamarBlue but without test agent at 570, and P2 isthe absorbance of the control wells with AlamarBlue but without the testagent at 600.% difference between=(O2×A1)−(O1×A2)×100treated and untreated cells(O2×P1)−(O1×P2)

The dose response data generated by these values was used to calculatethe half maximal inhibitory concentration (IC50) using a regressioncurve (3 or 4 parameter logistic sigmoidal curve or sigmoidal sigmoidcurve) in Sigma Plot, version 12.5. Statistical differences werecalculated using SPSS Statistics software, version 17.0, using theindependent t-test with an α of 0.05.

Prior to initiation of the studies using the anti-proliferative drugs,the cell density for the assay and AlamarBlue incubation time wasqualified. A range of cell concentrations was tested with repeatedabsorbance readings at established time points, then the resultinggraphs were used to determine the optimum cell plating density. As seenin FIG. 1 and FIG. 2, the cell density controlled the optimal incubationtime, with denser cells requiring less time to reach maximum reductionof AlamarBlue.

After examination of the curves generated by the experimental data shownin FIG. 1 and FIG. 2, we selected 5×10³ as the plating density. Thisconcentration performed well at every time point and fit within thelinear portion of the curve.

For the incubation time, we identified the time between 2 and 7 hours tobe ideal for the predetermined cell concentration, as the values fromthose incubation times fall within a linear curve and appeared lessnoisy than the longer incubations. During experiments with theanti-proliferative drugs, the plates were read at 2, 4 and 7 hours postaddition of AlamarBlue, in order to have the opportunity to capture thebest quality data. The 4-hour time point generally provided the datawith the more complete calorimetric development with lower variance.

In the human colorectal adenocarcinoma cell line HT29, there weredistinctly different dose responses for the two novel anti-proliferativedrugs in comparison to SN38 as shown in FIG. 3. SN38 showed a high levelof inhibition in this cell line throughout all concentrations tested,while compounds 2 (LGP001) and 3 (LGP002) had a gradual tapering ofproliferative inhibition that paralleled the decrease in concentration.The dose response curve was steeper in cells treated with compound 3(LGP002) in comparison to compound 2 (LGP001), showing lower %inhibition as the concentration of anti-proliferative decreased. At thehighest concentration tested, 100 μM, compounds 2 (LGP001) and 3(LGP002) were not significantly different from SN38 (p=0.984 andp=0.928, respectively). However, as the concentration ofanti-proliferative compound decreased, the differences in % inhibitionversus SN38 were significant. At 33.3 μM, both compounds 2 (LGP001) and3 (LGP002) showed high inhibition, above 54%, which was significantlydifferent from SN38 for both compounds 2 (LGP001) (p=0.030) and 3(LGP002) (p=0.003).

TABLE 1 IC50 of anti-proliferative drugs in HT29 cells. IC50 wascalculated using a 4 parameter sigmoidal logistic regression curve forcompounds 2 (LGP001) and 3 (LGP002), and a 3 parameter sigmoidallogistic regression curve for SN38. IC50 2 (LGP001) 5.6 3 (LGP002) 17.1SN38 0.2

The IC50 for all three anti-proliferative drugs in HT29 cells wascalculated and shown in Table 1. In agreement with the inhibitoryresponse seen at even the lowest tested dose, the IC50 of SN38 was thelowest of the three compounds, followed by 2 (LGP001) at 5.6 and finally3 (LGP002) at 17.1.

In the human colorectal adenocarcinoma cell line SW620, there were againdistinctly different dose responses for the two novel anti-proliferativedrugs in comparison to SN38 as shown in FIG. 4. SN38 had a high level ofinhibition in this cell line throughout all concentrations tested, over55%, while compounds 2 (LGP001) and 3 (LGP002) had a gradual tapering ofproliferative inhibition paralleling the decrease in concentration. Thedecrease in inhibitory effect was more pronounced in cells treated withcompound 3 (LGP002) as compared to compounds 2 (LGP001). These cellsalso appeared to be less sensitive to the anti-proliferative compoundsthan the other human cell line studied i.e., HT29, which had a lower %inhibition for compounds 2 (LGP001), 3 (LGP002) and SN38 at the lowestconcentration, although it was only statistically significant for SN38(p=0.025). In the SW620 cells, both compounds 2 (LGP001) and 3 (LGP002)were significantly different from SN38 in activity for allconcentrations tested, except for compound 2 (LGP001) at 33 μM(p=0.0369).

TABLE 2 IC50 of anti-proliferative drugs in SW620 cells. IC50 wascalculated with a 4 parameter sigmoidal logistic regression curve forcompounds 2 (LGP001) and SN38, and a 4 parameter sigmoidal sigmoidregression curve for compounds 2 (LGP001) and 3 (LGP002). IC50 2(LGP001) 1.9 3 (LGP002) 10.3 SN38 0.7

The IC50 for all three anti-proliferative drugs in SW620 cells wascalculated and shown in Table 2. Correlating to the inhibitory responseseen at even the lowest tested dose, the IC50 of SN38 was the lowest ofthe three compounds, followed by 2 (LGP001) at 1.9 and finally 3(LGP002) at 10.3.

In the murine colorectal carcinoma cell line CT26, there were againdistinctly different dose responses for the two novel anti-proliferativedrugs in comparison to SN38 as shown in FIG. 5. SN38 had the higherlevel of inhibition for this cell line throughout all concentrationstested, although this was generally lower than what was observed in thehuman colorectal cancer cell lines. Compounds 2 (LGP001) and 3 (LGP002)had a gradual tapering of proliferative inhibition paralleling thedecrease in concentration. The decrease in inhibitory effect was morepronounced in cells treated with compound 3 (LGP002) than compound 2(LGP001). These cells also appeared to be less sensitive to theanti-proliferative compounds than the human cell lines, having a lower %inhibition for compound 2 (LGP001) versus SW620 cells (p=0.000) and HT29cells (p=0.002) at the lowest concentration tested. This was alsoobserved with CT26 cells treated with compounds 3 (LGP002), which had alower % inhibition than SW620 cells (p=0.002) and HT29 cells (p=0.05) atthe lowest concentration tested. Similar to the results seen between thetwo human colorectal cancer cell lines, CT26 also had a significantlylower % inhibition when treated with SN38 versus SW620 cells (p=0.000)and HT29 cells (p=0.024) at the lowest concentration. Both compounds 2(LGP001) and 3 (LGP002) were significantly different from SN38 inactivity for all concentrations tested.

The IC₅₀ for all three anti-proliferative drugs in CT26 cells wascalculated and shown in Table 3. In contrast to the results seen in theother two cell lines, compound 3 (LGP002) had the lower IC50 (14.2)compared to compound 2 (LGP001) (18.0) in the CT26 cells. SN38 continuedto have the lowest IC₅₀ of the three compounds, at 0.6. SN38 continuedto have a consistently low IC₅₀ with the murine cell line that wascomparable to the IC₅₀ seen in the two human cell lines (0.6 for CT26versus 0.7 in the SW620 cells and 0.2 in the HT29 cells). Thus, in thesethree cell lines, SN38 provided a consistent control value forinhibitory capability of anti-proliferative compound.

TABLE 3 IC50 of anti-proliferative drugs in CT26 cells. IC50 wascalculated using a sigmoidal logistic 4 parameter regression curve forall compounds. IC50 2 (LGP001) 18.0 3 (LGP002) 14.2 SN38 0.6

The invention claimed is:
 1. A compound having the structure:


2. A pharmaceutical composition comprising the compound of claim 1 and avehicle.
 3. A method of treating colon cancer in a subject comprisingadministering to the subject in need thereof a therapeutically effectiveamount of the compound of claim 1.